| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Vanguard (LW) | A | NT | |
| 4 | McLaughlin | A | NT | |
| 5 | Lake Region | A | NT |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Santa Fe Catholic | A | NT | |
| 2 | Frostproof | A | 46.31 | |
| 3 | Bartow | A | DNS | Poinciana Relays |
| 4 | Haines City | A | 44.20 | |
| 5 | Fort Meade | A | 44.62 | Kowboy Invitational |
| 6 | Kathleen | A | 45.00 | |
| 7 | Discovery | A | 47.16 | Poinciana Relays |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Auburndale | A | 43.58 | |
| 2 | Lake Gibson | A | 42.94 | 42.94 Windermere Meet of Champions Classic 2023 |
| 3 | George Jenkins | A | 42.64 | Lakeland City meet @ George Jenkins 2023 |
| 4 | Victory Christian (LL) | A | DQ | Shorecrest Senior Night |
| 5 | Lake Wales | A | 42.44 | |
| 6 | Lakeland | A | 42.69 | Tohopekliga Tigers Invitational |
| 7 | Winter Haven | A | 43.51 | |
| 8 | Davenport | A | 43.83 | Poinciana Relays |